Two novel aspirin analogues show selective cytotoxicity in primary chronic lymphocytic leukaemia cells that is associated with dual inhibition of Rel A and COX-2.

OBJECTIVES: Non-steroidal anti-inflammatory drugs have been shown to induce apoptosis in primary B-cell chronic lymphocytic leukaemia (CLL) cells, but the molecular mechanisms that underpin this observation have not been fully elucidated. Here, we have analysed the effect two novel aspirin analogues, 2-hydroxy benzoate zinc (2HBZ) and 4-hydroxy benzoate zinc (4HBZ), on primary CLL samples. MATERIALS AND METHODS: Cytotoxic effects of 2HBZ and 4HBZ were analysed in primary CLL cells derived from 52 patients, and normal B- and T-lymphocytes. Mechanisms of action of these agents were also elucidated. RESULTS: Both analogues induced apoptosis in a dose-dependent and time-dependent manner. Apoptosis was associated with activation of caspase-3 that could be partially abrogated by the caspase-9 inhibitor (Z-LEHD.fmk). Importantly, both agents demonstrated preferential cytotoxicity in CLL cells when compared to normal B- and T-lymphocytes. In terms of their molecular mechanisms of action, 4HBZ and 2HBZ inhibited COX-2 transcription and protein expression and this was associated with upstream inhibition of transcription factor Rel A. Co-culture of CLL cells with CD40 ligand-expressing mouse fibroblasts significantly increased COX-2 expression and inhibited spontaneous apoptosis. Importantly, the most potent analogue, 4HBZ, overcame pro-survival effects of the co-culture system and significantly repressed COX-2. Finally, elevated COX-2 expression was associated with poor prognostic subsets and increased sensitivity to 4HBZ. CONCLUSIONS: Our results demonstrate therapeutic potential of 4HBZ and are consistent with a mechanism involving suppression of Rel A nuclear translocation and inhibition of COX-2 transcription.

The cyclin-dependent kinase inhibitor SNS-032 has single agent activity in AML cells and is highly synergistic with cytarabine.

SNS-032 (BMS-387032) is a selective cyclin-dependent kinase (CDK) inhibitor. In this study, we evaluated its effects on primary acute myeloid leukemia (AML) samples (n=87). In vitro exposure to SNS-032 for 48 h resulted in a mean LD(50) of 139±203 nM; Cytarabine (Ara-C) was more than 35 times less potent in the same cohort. SNS-032-induced a dose-dependent increase in annexin V staining and caspase-3 activation. At the molecular level, SNS-032 induced a marked dephosphorylation of serine 2 and 5 of RNA polymerase (RNA Pol) II and inhibited the expression of CDK2 and CDK9 and dephosphorylated CDK7. Furthermore, the combination of SNS-032 and Ara-C showed remarkable synergy that was associated with reduced mRNA levels of the antiapoptotic genes XIAP, BCL2 and MCL1. In conclusion, SNS-032 is effective as a single agent and in combination with Ara-C in primary AML blasts. Treatment with Ara-C alone significantly induced the transcription of the antiapoptotic genes BCL2 and XIAP. In contrast, the combination of SNS-032 and Ara-C suppressed the transcription of BCL2, XIAP and MCL1. Therefore, the combination of SNS-032 and Ara-C may increase the sensitivity of AML cells to the cytotoxic effects of Ara-C by inhibiting the transcription of antiapoptotic genes.

Measured and simulated performance of a ceramic micromechanical beam steering device at 94 GHz.

We report the first experimental demonstration of a transmission-mode micromechanical beam steering device for use in standoff terahertz imaging and spectroscopy. The device was constructed by laminating laser-cut 96% alumina sheets to form two plates with interlocking rectangular gratings of 762 microm period and was characterized at 94 GHz in a free-space measurement setup with an automated elevation scan. Plate tilts as great as 6 degrees deflected the transmitted beam by 6 degrees for the transverse electric (TE) polarization and by 4 degrees for the transverse magnetic polarization. Finite-difference time-domain simulations of the TE performance were in good agreement with the measurements.

Imprinted diffractive optics for terahertz radiation.

Terahertz diffractive optic elements have been fabricated in polypropylene by imprinting with a silicon master. A silicon master is created with eight phase levels for high diffraction efficiency and etched using inductively coupled plasma reactive ion etching. This technique enables the rapid replication of complex optical structures in a high transmission material. Excellent replication of multilevel high efficiency Fresnel lenses is shown. The resulting lenses were tested with a 2 THz quantum cascade laser. The signal strength at the focus was 70 times the base signal strength.

Characterization of T-ray binary lenses.

Multilevel phase-shift Fresnel diffractive zone plates fabricated on silicon wafers have been used as T-ray imaging lenses. The imaging results, including spatial and temporal distribution of T-rays measured at the focal planes in the frequency range from 0.5 to 1.5 THz, indicate that the performance of the diffractive terahertz (THz) lens is comparable with or better than that of conventional refractive THz lenses. The unique properties of the T-ray binary lens make it possible to fabricate excellent optics for narrow-band THz applications.